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DNA Helicases

DNA Helicases

DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...

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Related Experiment Video

Updated: Mar 18, 2026

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Published on: November 10, 2016

Helicase Assays.

Xin Wang¹, Jing Li², Jason Diaz³

¹Depatment of Molecular Genetics, Cleveland Clinic Lerner Research Institute, Cleveland, USA.

|July 9, 2016

Summary

This study details a new method for analyzing DNA helicase activity in vitro. The protocol uses the Merkel cell polyomavirus large T-antigen helicase, immobilized on IgG resin for efficient assessment.

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Last Updated: Mar 18, 2026

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Published on: November 10, 2016

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Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase

Single-Molecule Real-Time Visualization of DNA Unwinding by CMG Helicase

Published on: September 27, 2024

Area of Science:

Biochemistry
Molecular Biology
Enzymology

Background:

Helicases are essential motor proteins that unwind nucleic acid strands.
These enzymes play critical roles in fundamental cellular processes like DNA replication and repair.
Understanding helicase function requires robust in vitro assay methods.

Purpose of the Study:

To develop and present a protocol for analyzing DNA helicase activity in vitro.
To demonstrate the utility of a specific helicase, Merkel cell polyomavirus large T-antigen, in this assay.

Main Methods:

Expression of Merkel cell polyomavirus large T-antigen in HEK293 cells.
Immobilization of the purified helicase protein onto an IgG resin.
Performing helicase activity assays with the immobilized protein.

Main Results:

Successful expression and purification of the target DNA helicase.
Demonstration of a functional in vitro helicase assay using immobilized enzyme.
The protocol allows for the analysis of helicase activity under specific conditions.

Conclusions:

The presented protocol provides a reliable method for studying DNA helicase activity.
Immobilization of helicases on resin facilitates in vitro biochemical analysis.
This method can be adapted for various helicase proteins and research questions.